Fig. 2

Effect of Smyd3 knockdown in zebrafish embryos by Smyd3-MO or Smyd3-SB-MO.

(A, B, C, and D) Suppression of smyd3 was examined at 10 hpf in embryos injected with smyd3-EGFP mRNA alone (A), smyd3-EGFP mRNA and Smyd3-mis-MO (B), and smyd3-EGFP mRNA and Smyd3-MO (C). Signals of EGFP were examined in the fluorescent macroscope (lower panel). The frequencies of EGFP-positive embryos were (A) 78.7%±2.3, (B) 83.7%±1.5, and (C) 17.7%±3.4. Data are shown as means±SEM. (D) Effect of Smyd3-SB-MO on smyd3 transcripts. The wild-type transcripts were detected as a band at 465 bp and the aberrant transcripts at 401 bp. The lower panel shows the expression of ef1α as a control. (E and F) Phenotype of embryos injected with Smyd3-MO (E) or Smyd3-mis-MO (F) at 72 hpf. The pericardial edema (arrow head) and curved trunk (arrow) were observed in Smyd3 morphants (E). (G, H, I, and J) Morphological classification of heart defect. The degree of cardiac defect in the morphants was classified into three grades at 48 hpf. Grade1: Heart shows abnormality with mild looping defect and pericardial edema (H); Grade2: Heart shows abnormality with moderate looping and defect and pericardial edema (I); Grade3: Heart shows abnormality with string-like heart with severe pericardial edema (J). Normal: Normal heart (G). Embryos are shown in lateral view.